Sie sind auf Seite 1von 14

Infection Strategies of Botrytis cinerea

Jan A.L. van Kan


Wageningen University, Laboratory of Phytopathology, Binnenhaven 5
6709 PD Wageningen
The Netherlands
Keywords: grey mould, post-harvest damage, molecular genetics, virulence genes.
Abstract
Botrytis cinerea is a ubiquitous filamentous fungal pathogen of a wide range
of plant species. The fungus is able to infect all aerial parts of its host plants to a
certain extent. Infection may cause enormous damage both during plant growth and
in the post-harvest phase (during cold storage or transport). B. cinerea is a major
cause of economic loss in the production chain of cut flowers, bulb flowers and pot
plants. Molecular-genetic studies performed over the past decade have provided a
wealth of novel insights into the infection mechanisms utilised by the pathogen.
Fungal genes were identified that are important for successful infection by B.
cinerea. Such knowledge provides perspectives for designing novel, rational plant
protection strategies that effectively counteract important fungal virulence factors.
In this review I will divide the infection process into different stages and discuss the
role of various fungal enzymes and metabolites in the individual stages. Finally some
perspectives are addressed for novel control strategies that may reduce and/or delay
the damage inflicted by B. cinerea infection.
INTRODUCTION
Botrytis cinerea Persoon: Fries (teleomorph Botryotinia fuckeliana, also known as
grey mould fungus) causes serious pre- and post-harvest diseases in at least 200 plant
species (Jarvis, 1977), including agronomically important crops and harvested
commodities, such as grapevine, tomato, strawberry, cucumber, bulb flowers, cut flowers
and ornamental plants. The pathogen is a necrotroph, inducing host cell death resulting in
serious damage to plant tissues culminating in rot of the plant or the harvested product.
Reviews have been published on the infection mechanisms of B. cinerea, with emphasis
on microscopic and biochemical studies (Staples and Mayer, 1995) or molecular genetics
(Prins et al., 2000b). The application of molecular-genetic tools such as transformation
(Hamada et al., 1994), differential gene expression analysis (Benito et al., 1996), gene
cloning (van der Vlugt-Bergmans et al., 1997a,b) and targeted mutagenesis (van Kan et
al., 1997) has led to novel insight in the structure of B. cinerea genes and their role in the
infection process. Here our current understanding of the mechanisms that B. cinerea
exploits to infect its host plants will be summarised.
DISEASE CYCLE
For the purpose of simplification, different stages are distinguished in the disease
cycle of B. cinerea (Fig. 1). The disease cycle starts with a conidium landing on the host
surface. B. cinerea conidia are ubiquitous in the air and can be transported by wind over
long distances before infecting the next host (Jarvis, 1977). Following attachment, the
conidium germinates on the host surface under moist conditions and produces a germ tube
that develops into an appressorium that penetrates the host surface. The underlying cells
are killed and the fungus establishes a primary lesion, in which necrosis and host defense
responses may occur. In some cases this is the onset of a period of quiescence of an
undefined length, in which fungal outgrowth is restricted (reviewed by Prusky, 1996).
Quiescence is especially an important feature in host species infected at the flowering
stage (e.g. strawberry). At a certain stage, the defense barriers are breached and the
fungus starts a vigorous outgrowth, resulting in rapid maceration of plant tissue, on which
the fungus eventually sporulates to produce inoculum for the next infection. Under
Proc. VIIIth IS Postharvest Phys. Ornamentals
Eds. N. Marissen et al.
Acta Hort. 669, ISHS 2005

77

optimal conditions, an infection cycle may be completed in as little as 3-4 days,


depending on the type of host tissue attacked. In the following paragraphs I will discuss
the different stages in the disease cycle and the role that enzymes and metabolites (mainly
of fungal origin) play in these stages. Given the wide host range of the fungus, not all
stages or processes may occur in every infection.
Attachment of Conidia
Attachment is thought to be mediated by physical surface interactions on the plant
cuticle. Two steps were distinguished in the attachment to host tissue. The first stage,
preceding the hydration of conidia, typically involves weak adhesive forces resulting
from hydrophobic interactions between host and conidial surfaces (Doss et al., 1993).
Stronger binding occurs in the second stage (Doss et al., 1995), several hours after
inoculation, when conidia have germinated. The tips of germ tubes are covered with
fibrillar-like extracellular matrix material (Doss et al., 1995; Cole et al., 1996; see also
Fig. 2 in Prins et al., 2000b), consisting of carbohydrates and proteins (Doss, 1999). The
matrix contains fungal enzymes (Gil-Ad et al., 2001). It may act as an adhesive on the
host surface (Doss et al. 1995) and protect hyphae from dehydration and defense
mechanisms of the host.
Germination
Several factors influence the germination of a conidium. Free surface water or
high relative humidity (>93% RH) is essential to germinate and penetrate the host
epidermis (Williamson et al., 1995). When dry conidia are inoculated on plant surfaces
and incubated in the absence of free surface water, the emerging germ tube usually
remains short before it penetrates the surface (Salinas and Verhoeff, 1995; Williamson et
al., 1995; Cole et al., 1996). Inoculation with conidia in an aqueous suspension requires
the addition of nutrients (Harper et al., 1981; van den Heuvel, 1981), which might mimic
the situation in a wound on the plant epidermis, from which nutrients leach. A highly
efficient germination and synchronous infection of tomato leaves was obtained, when
conidia were pre-incubated for 2-4 hours in liquid medium supplemented with phosphate
and sugar, prior to inoculation (Benito et al., 1998).
Gaseous compounds may stimulate germination. Elad and Volpin (1988) found a
correlation between the level of ethylene production by rose cultivars and the severity of
grey mould symptoms. Stimulation of grey mould development by ethylene was also
found in strawberry, tomato, cucumber and pepper (e.g. Elad and Volpin, 1988; McNicol
et al., 1989). This observation is usually ascribed to host tissue senescence that coincides
with ethylene production. The influence of ethylene on B. cinerea itself has not been
studied in detail. Analogous to Colletotrichum gloeosporioides (Flaishman and
Kolattukudy, 1994), germination of B. cinerea seems to be influenced by ethylene.
Germination of conidia on a hydrophobic surface was stimulated by exogenous ethylene,
but the germ tube length was unaffected (Kepczynski and Kepczynska, 1977).
Application of 2,5-norbornadiene, a competitive inhibitor of ethylene perception in plants,
inhibited germination in a reversible manner (Kepczynska, 1993). In a hydrophilic
environment, however, ethylene stimulated germ tube elongation without affecting the
percentage of germination (Barkai-Golan et al., 1989). Ethylene produced by the plant
during tissue senescence or fruit ripening might function as a signal for the conidia on the
(hydrophobic) plant surface to germinate and initiate the infection. Germ tube elongation
might subsequently be stimulated by ethylene in the more hydrophilic environment of the
invaded plant tissue. Thus, ethylene might favour grey mould development by weakening
the host, as well as by stimulating germination of B. cinerea conidia and outgrowth of
hyphae. Molecular and biochemical approaches are required to further elucidate the
effects of ethylene on B. cinerea.
Differentiation of Infection Structures on the Host Surface
B. cinerea forms appressoria during penetration, albeit not the highly organized
78

appressoria that are typical for many plant pathogenic fungi (reviewed by Mendgen et al.,
1996). Several authors observed the swelling of hyphal tips of germ tubes and interpreted
these as an appressorium-like structure (Akutsu et al., 1981; van den Heuvel and
Waterreus, 1983; Cole et al., 1996). Recent microscopic and histochemical studies
(Tenberge et al., 2002) and gene function analysis (Gourgues et al., 2003) indicate that
these structures act as functional appressoria. The swelling of the hyphal tip may be the
consequence of a rise in osmotic value in the hyphal tip, resulting in water absorption. In
the absence of a rigid layer in the outer wall, swelling cannot result in an equally high
turgor as in the appressoria of Magnaporthe grisea (Howard et al., 1991; de Jong et al.,
1997). The extracellular matrix may contribute to the swelling by retaining water, as its
major polysaccharide component (cinerean), is extremely hygroscopic.
Penetration of the Host Surface
Invasion of host tissue can be achieved by active penetration or passive ingress. B.
cinerea is an opportunist that can initiate infection at wound sites, or at sites previously
infected by other pathogens. B. cinerea can also enter the substomatal cavity via an open
stoma. Nevertheless the pathogen is perfectly able to penetrate intact host surfaces. Only
direct penetration of the epidermal surface is discussed in this paragraph. For reasons of
simplicity the penetration of dead or wounded tissue, as well as via stomata, is regarded
as an expansion rather then a penetration process, and is dealt with in later paragraphs.
The cuticle consists of cutin, a polyester of hydroxylated and epoxidized C16- and
C18-fatty acids, covered with wax. Physical damage or brute mechanical penetration of
the cuticle by B. cinerea is not usually observed (Williamson et al., 1995; Cole et al.,
1996). Hence, it was considered that enzymatic (cutinolytic) activity is required for
penetrating intact host surfaces (Salinas and Verhoeff, 1995; van der Vlugt-Bergmans,
1997a). Salinas (1992) raised monoclonal antibodies against a 18 kDa cutinase.
Application of the antibody to gerbera flowers prior to inoculation reduced the number of
lesions formed. The gene encoding this cutinase enzyme was cloned (van der VlugtBergmans et al., 1997a) and a gene replacement mutant was made that was entirely
devoid of this cutinase activity (van Kan et al., 1997). The mutant was equally virulent as
the wild type isolate, both on gerbera flowers and tomato fruits and the fungus remained
able to penetrate intact cuticle surfaces (van Kan et al., 1997). Although the observations
of Salinas (1992) remain to be explained, it can be concluded that this 18 kDa cutinase is
not essential in penetration.
A different enzyme that may mediate host penetration is a 60 kDa lipase
(Commnil et al., 1995), inducible by apple cutin (Commnil et al., 1998) as well as
grape berry cuticle components (Commnil et al., 1999). The lipase possesses cutinolytic
activity although with clearly distinct kinetic properties than the 'typical' cutinase
mentioned above (Commnil et al., 1998). When polyclonal antibodies raised against this
lipase were applied prior to inoculation with B. cinerea conidia, germ tubes were
prevented from penetrating the cuticle. The antibodies did not affect germination
(Commnil et al., 1998). The protein has been partially sequenced (Commenil et al.,
1999). Constructing a targeted lipase-deficient mutant and determining its virulence
should assess the role of the lipase in host tissue penetration.
Killing the Host
B. cinerea kills host cells before they are invaded by hyphae (Clark and Lorbeer,
1976). Invasion of plant tissue by B. cinerea triggers nuclear condensation and plant
membrane damage, indicators for programmed cell death, in a ring of cells around the
hyphae (Govrin and Levine, 2000). These results imply that diffusible factors have a
direct or indirect phytotoxic activity. These factors may be proteins or low molecular
weight compounds secreted by the fungus into its environment. The induction of
programmed cell death facilitates B. cinerea invasion and may in fact be essential for
successful infection (Govrin and Levine, 2000).
1. Toxins. Culture filtrates of B. cinerea may induce toxic effects when applied to plant
79

tissue (Rebordinos et al., 1996). Phytotoxic compounds were identified as botcinolide, a


highly substituted lactone (Cutler et al., 1993) and botrydial, a tricyclic sesquiterpene
(Colmenares et al., 2002). The observation that both secondary metabolites were only
secreted by B. cinerea in medium with high glucose levels initially raised doubts about
their physiological relevance in planta. Recent analytical chemistry studies have,
however, demonstrated that botrydial accumulates in infected tissue at physiologically
relevant concentrations (Deighton et al., 2001; Muckenschnabel et al., 2003). The
production of botrydial may be an important factor in the infection of (some) host plants,
but it needs to be evaluated by constructing mutants in the botrydial synthetic pathway.
Resolution of this pathway is in progress (Duran et al., 2001) but the relevant botrydial
biosynthetic genes have not yet been identified and mutants are not available.
2. Oxalic Acid. Secretion of oxalic acid (OA) occurs in fungi from various taxonomic
classes (reviewed by Dutton and Evans, 1996). A key role for OA in pathogenesis has
been postulated for Sclerotinia sclerotiorum (Godoy et al., 1990), a close relative of B.
cinerea. Mutants of S. sclerotiorum, deficient in OA production, were unable to infect
Arabidopsis plants (Dickman and Mitra, 1992) and the deficiency could be restored by
supplementing the inoculum with OA. B. cinerea produces OA both in vitro (Gentile,
1954; Germeier et al., 1994) and in planta (Verhoeff et al., 1988). OA may form calcium
oxalate crystals within the host tissue (see Figure 3 in Prins et al., 2000b). The sizes of
lesions induced by several strains of B. cinerea on grapevine and bean leaves correlated
with the amount of OA that these strains secreted in vitro (Germeier et al., 1994). It
remains unclear whether the amounts of OA produced in planta are sufficient to cause
host cells to collapse. OA may in fact be a co-factor in pathogenesis rather than the
primary phytotoxic agent. Ten Have et al. (2002) postulated that OA might act in synergy
with endoPGs during tissue maceration. Fungal endoPGs have an activity optimum at low
pH and may therefore be stimulated by the simultaneous secretion of OA. Moreover, OA
may stimulate pectin degradation resulting from endoPG action by sequestering the Ca2+
ions from (intact or partially hydrolyzed) Ca-pectates in the cell walls. The removal of
Ca2+ ions disturbs intermolecular interactions between pectic polymers and disrupts the
integrity of the pectic backbone structure. Consequently, the pectic structure absorbs
water and swells, as described by Mansfield and Richardson (1981).
3. Induction of Reactive Oxygen Species. Recent studies have focussed on Reactive
Oxygen Species (ROS) production in relation to B. cinerea pathogenicity. ROS is the
joint term for the superoxide anion, hydroxyl radical and hydrogen peroxide. The level of
H2O2 released from bean leaf discs inoculated with different B. cinerea isolates,
correlated with the aggressiveness of the isolate on such leaf tissue (Tiedemann, 1997).
The sensitivity of genotypes of bean (Phaseolus vulgaris) to oxidative stress is correlated
to their susceptibility to B. cinerea (Tiedemann, 1997). In B. cinerea-infected Arabidopsis
thaliana leaves, H2O2 was detected in the apoplastic space several cell layers away from
the fungal hyphae (Govrin and Levine, 2000). Enormous perturbances in redox status are
observed at the host-fungal interface, even in plant tissue at some distance from the
infection front (Muckenschnabel et al., 2001a,b; 2002). Also lipid peroxidation was
observed (Weigend and Lyr, 1996; Muckenschnabel et al., 2001a; 2002). Antioxidants
reduce grey mould disease development (Elad, 1992; Tiedemann, 1997).
Generation of ROS occurs at the host-fungal interface. Fungal as well as host
enzymes presumably contribute to the process (Schouten et al., 2002a; see also Fig. 4 in
Prins et al., 2000b). Oligogalacturonides released from the plant cell wall by pectinases of
B. cinerea are potential elicitors of an oxidative burst (Legendre et al., 1993), presumably
mediated by a plasmamembrane-bound NADPH oxidase, inducible by fungal elicitors
and requiring extracellular Ca2+ (Schwacke and Hager, 1992). Indeed Schouten et al.
(2002a) demonstrated the production of H2O2 in the plasmamembrane of host cells
adjacent to a fungal hypha. Infiltration of the NADPH oxidase inhibitor DPI into leaves of
Arabidopsis thaliana prior to inoculation with B. cinerea resulted in a reduction of ROS
production and slower colonization of host tissue by the fungus (Govrin and Levine,
2000). The precise mechanism by which B. cinerea induces oxidative stress in its hosts is
80

still unclear. Fungal extracellular sugar oxidases (Liu et al., 1998; Edlich et al., 1989) or
superoxide dismutase (Tenberge et al., 2002) may be responsible for generating the H2O2.
The identification of the fungal enzymes that are important for H2O2 accumulation at the
host-fungus interface awaits molecular-genetic studies with targeted mutants.
Formation of Primary Lesions, Defense Responses in the Host
Host surface penetration and the rupture of plant cell walls by enzymes of B.
cinerea triggers a cascade of processes in the fungus as well as the host. This paragraph
deals with defense responses at the host-fungus interface and their impact on the progress
of infection.
1. Induction of Necrosis. The initial establishment of primary necrotic lesions coincides
with (and is in fact the result of) host defense activation in the neighbouring tissue in
response to the death of an invaded cell. It is as yet unclear whether cell death caused by a
necrotroph, such as B. cinerea, is equivalent to cell death during a hypersensitive
response (HR) to a biotrophic pathogen (reviewed by Lamb and Dixon, 1997). Govrin
and Levine (2000) showed that an oxidative burst occurs in plant tissue several cell layers
away from the fungal hyphae. Cytological staining provided evidence for rapid nuclear
condensation and irreversible membrane damage, indicative of a programmed cell death
process (Govrin and Levine, 2000). Largely the same defense responses are activated
during an infection by B. cinerea as during HR to avirulent races of a biotrophic
pathogen: lignification (Maule and Ride, 1976; Heale and Sharman, 1977), biosynthesis
of phytoalexins (e.g. Bennett et al., 1994) and accumulation of PR proteins (e.g. Benito et
al., 1998; Diaz et al., 2002). The total spectrum of defense responses results in a primary
necrotic lesion in which the fungus is effectively restricted. Depending on the type of host
tissue and yet unidentified physiological aspects of the host, the lesions enter a lag phase
in which they do not expand. A proportion of the primary lesions eventually develops into
aggressive, expanding lesions (van den Heuvel, 1981; De Meyer and Hfte, 1997; Benito
et al., 1998; Diaz et al., 2002). In the non-expanding lesions the fungus is not killed, since
viable fungal mycelium could be recovered from all lesions (Benito and van Kan,
unpublished results). Thus, an active defense contributes to (temporarily) restricting the
fungus within the primary lesions, giving rise to a period of quiescence.
2. Quiescence. In some tissues, B. cinerea causes long-lasting quiescent infections
(reviewed by Prusky, 1996), in which no symptoms are discernible at first. Prominent
examples are described in soft fruit such as strawberry, raspberry and grape. In these
hosts, B. cinerea predominantly infects the host flowers and resides quiescent in the
developing fruit tissue, often for several weeks. Fungal growth resumes at the onset of
fruit ripening. It has been postulated that high levels of fungitoxic or fungistatic
compounds (phytoalexins) in immature fruits contribute to grey mould quiescence. The
level of these compounds decreases during the ripening process concomitant with fungal
outgrowth. Attempts have been undertaken to increase the levels of antifungal compounds
or to prevent their degradation during ripening. The level of the stilbene phytoalexin
resveratrol in grapes is correlated with grey mould resistance (Langcake, 1981; Bavaresco
et al., 1997). The effect of over-expressing stilbene synthase genes from Vitis in
transgenic plants on resistance towards B. cinerea was evaluated. A significant, partial
resistance was obtained in tobacco (Hain et al., 1993) but not in tomato (Thomzik et al.,
1997).
Besides phytoalexins, immature fruits usually contain high levels of proteinaceous
inhibitors of fungal cell wall degrading enzymes, the PolyGalacturonase Inhibiting
Proteins (PGIPs) and their level decreases during ripening (De Lorenzo et al., 2001). In
view of the significant role that polygalacturonases play in the infection (see below),
efforts to produce transgenic plants overexpressing PGIPs have been undertaken to obtain
resistance towards B. cinerea. Indeed high constitutive expression of a heterologous PGIP
gene in tomato and an endogenous PGIP gene in Arabidopsis resulted in an increased
resistance to B. cinerea (Powell et al., 2000; Ferrari et al., 2003). One of the
considerations in this strategy is that PGIPs have a differential activity towards individual
81

fungal endoPGs (Desiderio et al., 1997; De Lorenzo et al., 2001). This makes it relevant
to chose PGIPs that are most potent against the B. cinerea endoPG isozymes that are
important in virulence (Ten Have et al., 2002).
Evasion of Chemical Defense
Pathogenic fungi have developed mechanisms to overcome deleterious effect of
preformed (phytoanticipins) or induced (phytoalexins) plant defense compounds. One
such mechanism involves an energy-dependent secretion by ABC-transporters that
confers some degree of tolerance to the fungitoxic effects of such compounds (de Waard,
1997). About a dozen ABC transporters have been cloned from B. cinerea (de Waard,
unpublished results) and their role in resistance towards plant defense compounds and
virulence on various hosts is being studied (Schoonbeek et al., 2001; 2002).
The major strategy of plant pathogenic fungi to deal with antifungal plant
compounds is an enzymatic detoxification (reviewed by Osbourn et al., 1998). Because of
its broad host range, B. cinerea encounters a wide spectrum of antimicrobial compounds,
depending on the host species that it attempts to infect. The ability of B. cinerea to
degrade or detoxify phytoalexins was already intensively studied two decades ago
(reviewed by Mansfield, 1980). The best studied example for phytoalexin detoxification
by B. cinerea is the detoxification of the Vitis phytoalexins pterostilbene and resveratrol.
The ability of fungal isolates to detoxify these phytoalexins was correlated with their
virulence (Sbaghi et al., 1996). Conversely, the resistance level of Vitis genotypes against
grey mould is correlated with their phytoalexin content (Langcake, 1981; Jeandet et al.,
1992). B. cinerea produces a substrate-specific laccase (stilbene oxidase) that is able to
oxidize both compounds to non-toxic derivatives (Pezet et al., 1991). The enzyme was
purified and characterized (Pezet, 1998) and the structure of stilbene degradation products
elucidated (Breuil et al., 1998). Schouten et al. (2002b) cloned a laccase gene responsible
for resveratrol conversion and generated targeted mutants that lost the ability to convert
resveratrol. The mutants did not show reduced virulence on Vitis or any other host tested
(Schouten et al., 2002b), indicating resveratrol conversion is not essential for virulence.
B. cinerea is also able to detoxify preformed antimicrobial compounds such as the
tomato saponin -tomatine (Verhoeff and Liem, 1975). Quidde et al. (1998) showed that
-tomatine is only partly deglycosylated by removal of the terminal xylose, yielding 1tomatine which is less toxic than -tomatine. A field survey showed that B. cinerea
isolates from various host plants and geographic origin all possessed tomatinase activity,
with one exception. Interestingly, the strain lacking tomatinase activity was highly
sensitive to -tomatine, completely non-pathogenic on tomato, yet highly aggressive on
Phaseolus vulgaris (Quidde et al., 1998). These data suggest that the ability to detoxify tomatine is correlated with virulence of B. cinerea on tomato. However, this strain might
have further defects that cause the specific loss of virulence on tomato; the cloning and
targeted mutagenesis of the tomatinase gene in a wild type strain will be necessary for
final evaluation of the role of tomatinase in the infection of tomato.
The oxidative burst at the host-pathogen interface imposes stress on the host as
well as the pathogen (Schouten et al., 2002a). B. cinerea is able to cope with external
oxidative stress in order to survive in the necrotic tissue. Successful detoxification of
H2O2 is presumably mediated by an extracellular catalase (Schouten et al., 2002a) with a
Glutathione S-Transferase functioning as intracellular back-up (Prins et al., 2000a;
Schouten et al., 2002a). Targeted mutagenesis of the extracellular catalase gene did not
negatively affect the survival of the mutant within the oxidative environment of a necrotic
lesion. Virulence of the mutant on several hosts was indistinguishable from that of the
wild type (Schouten et al., 2002a).
Disease Expansion and Tissue Maceration
B. cinerea must be able to macerate plant tissue and convert it into fungal
biomass. The initial step in expansion of primary lesions is presumably the killing of
neighbouring cells by mechanisms similar to the ones described above. In order to grow
82

from the primary lesion into neighbouring tissue, B. cinerea must actively degrade plant
cells. The major barrier that the pathogen encounters is the host cell wall. Cell wall
degradation facilitates the entry of the pathogen and it provides nutrients for growth
(reviewed by Ten Have et al., 2002).
Microscopic studies showed that after penetration of the cuticle, hyphae of B.
cinerea frequently invade the anticlinal wall between two epidermal cells. The
concomitant swelling of the epidermal cell wall (Mansfield and Richardson, 1981) is
indicative for the degradation of pectin in the matrix of the epidermal wall, resulting in
water absorption. Biochemical evidence suggested that pectinases are involved in primary
infection. B. cinerea possesses a set of cell wall degrading enzymes (CWDEs) including
pectin lyase (Movahedi and Heale, 1990), pectin methylesterase (Reignault et al., 1994;
Valette-Collet and Boccara, 2003), exo- and endo-polygalacturonase (Johnston and
Williamson, 1992) and cellulase (Barkai-Golan et al., 1988). B. cinerea genes have been
cloned that encode CWDEs: pectin and pectate lyase (Mulder, unpublished),
rhamnogalacturonan-hydrolase (Chen et al., 1997) six endoPGs (Ten Have et al., 1998,
Wubben et al., 1999), pectin methylesterase (Valette-Collet et al., 2003) and cellulases
(van Kan et al., unpublished). The endoPG genes, denoted Bcpg1-6, constitute a well
studied and, most probably, complete gene family. The expression patterns of the
individual endoPG genes in planta depend on the host species that is infected, on the stage
of the infection, as well as on the external conditions during which infection occurs (Ten
Have et al., 2001).
Targeted deletion mutants were made in Bcpg1 by gene replacement. The lesion
expansion rate of such mutants was reduced by about 25% as compared to the wild type
(Ten Have et al., 1998). The BcPG1 protein might be important in facilitating
intercellular growth at the periphery of the invading hyphae. Another explanation for the
reduction in virulence may be purely nutritional. The absence of BcPG1 reduces the
release of pectin degradation products that serve as nutrients, hence resulting in slower
growth of the fungus through the tissue. Mutagenesis of four additional endoPG genes is
in progress (van Kan et al., unpublished results). The distinction between a pathogenic
function and a nutritional function of B. cinerea endoPGs (Ten Have et al., 2002) is
difficult to make.
SUMMARY AND CONCLUDING REMARKS
Molecular-genetic tools are available to validate microscopic and biochemical
observations of the infection strategy of B. cinerea. Dozens of genes have been cloned
and their expression in planta or in vitro studied. The role(s) of individual genes in the
infection process can be analysed by targeted mutagenesis and studying the behaviour of
mutants on various hosts. This overview demonstrates that the pathogen is versatile and
uses a combination of factors during pathogenesis. We are beginning to understand the
roles of various factors in the different stages of the disease cycle and the ways in which
some of these factors interact. There seems to be a delicate balance between the attack
mechanisms of the fungus and the defense of the host. Such knowledge will contribute to
new, rational disease control strategies. The future challenge lies in the design of methods
that alter the balance in the interaction in favor of the host. Such a strategy will likely
consist of a sophisticated combination of biological control agents, appropriate (partially)
resistant plant genotypes and chemicals that either enhance the plant defense response or
interfere with crucial steps in the infection process.
ACKNOWLEDGEMENTS
Several research projects in my group are supported by the Technology
Foundation (STW), subsidized by the Ministry of Economic Affairs and The Netherlands
Organization for Scientific Research (NWO). I am grateful to Jos Raaijmakers for critical
reading of the manuscript.

83

Literature Cited
Akutsu, K., Kobayashi, Y., Matsuzawa, Y., Watanabe, T., Ko, K. and Misato, T. 1981.
Morphological studies on infection process of cucumber leaves by conidia of Botrytis
cinerea stimulated with various purine-related compounds. Ann. Phytopathol. Soc.
Japan 47: 234-243.
Barkai-Golan, R., Lavy Meir, G. and Kopeliovitch, E. 1988. Pectolytic and cellulolytic
activity of Botrytis cinerea Pers. related to infection of non-ripening tomato mutants.
J. Phytopathol. 123: 174-183.
Barkai-Golan, R., Lavy Meir, G. and Kopeliovitch, E. 1989. Stimulation of fruit ethylene
production by wounding and by Botrytis cinerea and Geotrichum candidum infection
in normal and non-ripening tomatoes. J. Phytopathol. 125: 148-156.
Bavaresco, L., Petegolli, D., Cantu, E., Fregoni, M., Chiusa, G. and Trevisan, M. 1997.
Elicitation and accumulation of stilbene phytoalexins in grapevine berries infected by
Botrytis cinerea. Vitis 36: 77-83.
Benito, E.P., Prins, T.W. and Van Kan, J.A.L.. 1996. Application of differential display
RT-PCR to the analysis of gene expression in a plant fungus interaction. Plant Mol.
Biol. 32: 947-957.
Benito, E.P., Ten Have, A., Van 't Klooster, J.W. and Van Kan, J.A.L. 1998. Fungal and
plant gene expression during synchronized infection of tomato leaves by Botrytis
cinerea. European J. Plant Pathol. 104: 207-220.
Bennett, M.H., Gallagher, M.D.S., Bestwick, C.S., Rossiter, J.T. and Mansfield, J.W.
1994. The phytoalexin response of lettuce to challenge by Botrytis cinerea, Bremia
lactucae and Pseudomonas syringae pv. phaseolicola. Physiol. Mol. Plant Pathol. 44:
321-333.
Breuil, A.C., Adrian, M., Pirio, N., Meunier, P., Bessis, R. and Jeandet, P. (1998)
Metabolism of stilbene phytoalexins by Botrytis cinerea: I. Characterization of a
resveratrol dehydrodimer. Tetrahedron Letters 39: 537-540.
Chen, H.J., Smith, D.L., Starrett, D.A., Zhou, D.B., Tucker, M.L., Solomos, T. and Gross,
K.C. 1997. Cloning and characterisation of a rhamnogalacturonan hydrolase gene
from Botrytis cinerea. Biochem. Mol. Biol. Int. 43: 823-838.
Clark, C.A. and Lorbeer, J.W. 1976. Comparative histopathology of Botrytis squamosa
and B. cinerea on onion leaves. Phytopathology 66: 1279-1289.
Cole, L., Dewey, F.M. and Hawes, C.R. 1996. Infection mechanisms of Botrytis species:
Pre-penetration and pre-infection processes of dry and wet conidia. Mycol. Res. 100:
277-286.
Colmenares, A.J., Aleu, J., Duran, R.P., Collado, I.G. and Hernandez, R.G. 2002. The
putative role of botrydial and related metabolites in the infection mechanism of
Botrytis cinerea. J. Chem. Ecol. 28: 997-1005.
Commnil, P., Belingheri, L., Sancholle, M. and Dehorter, B. 1995. Purification and
properties of an extracellular lipase from the fungus Botrytis cinerea. Lipids 30: 351356.
Commnil, P., Belingheri, L. and Dehorter, B. 1998. Antilipase antibodies prevent
infection of tomato leaves by Botrytis cinerea. Physiol. Mol. Plant Pathol. 52: 1-14.
Commnil, P., Belingheri, L., Bauw, G. and Dehorter, B. 1999. Molecular
characterization of a lipase induced in Botrytis cinerea by components of grape berry
cuticle. Physiol. Mol. Plant Pathol. 55: 37-43.
Cutler, H.G., Jacyno, J.M., Harwood, J.S., Dulik, D., Goodrich, P.D. and Roberts, R.G.
1993. Botcinolide: a biologically active natural product from Botrytis cinerea. Biosci.,
Biotechnol. Biochem. 57: 980-1982.
De Meyer, G. and Hfte, M. 1997. Salicylic acid produced by the rhizobacterium
Pseudomonas aeruginosa 7NSK2 induces resistance to leaf infection by Botrytis
cinerea on bean. Phytopathology 87: 588-593.
Deighton, N., Muckenschnabel, I., Colmenares, A.J., Collado, I.G. and Williamson, B.
2001. Botrydial is produced in plant tissues infected by Botrytis cinerea.
Phytochemistry 57: 689-692.
84

de Jong, J.C., McCormack, B.J., Smirnoff, N. and Talbot, N.J. 1997. Glycerol generates
turgor in rice blast. Nature 389: 244-245.
De Lorenzo, G., D'Ovidio, R. and Cervone, F. 2001. The role of polygalacturonaseinhibiting proteins (PGIPs) in defense against pathogenic fungi. Annual Rev.
Phytopathol. 39: 313-335.
Desiderio, A., Aracri, B., Leckie, F., Mattei, B., Salvi, G., Tigelaar, H., Van Roekel,
J.S.C., Baulcombe, D.C., Melchers, L.S. and De Lorenzo, G. 1997.
Polygalacturonase-inhibiting proteins (PGIPs) with different specificities are
expressed in Phaseolus vulgaris. Mol. Plant-Microbe Interact. 10: 852-860.
de Waard, M.A. 1997. Significance of ABC transporters in fungicide sensitivity and
resistance. Pesticide Science 51: 271-275.
Diaz, J., ten Have, A. and van Kan, J.A.L. 2002. The role of ethylene and wound
signaling in resistance of tomato to Botrytis cinerea. Plant Physiol. 129: 1341-1351.
Dickman, M.B. and Mitra, A. 1992. Arabidopsis thaliana as a model for studying
Sclerotinia sclerotiorum pathogenesis. Physiol. Mol. Plant Pathol. 41: 255-263.
Doss, R.P. 1999. Composition and enzymatic activity of the extracellular matrix secreted
by germlings of Botrytis cinerea. Appl. Environm. Microbiol. 65: 404-408.
Doss, R.P., Potter, S.W., Chastagner, G.A. and Christian, J.K. 1993. Adhesion of
nongerminated Botrytis cinerea conidia to several substrata. Appl. Environm.
Microbiol. 59: 1786-1791.
Doss, R.P., Potter, S.W., Soeldner, A.H., Christian, J.K. and Fukunaga, L.E. 1995.
Adhesion of germlings of Botrytis cinerea. Appl. Environm. Microbiol. 61: 260-265.
Dutton, M.V. and Evans, C.S. 1996. Oxalate production by fungi: its role in pathogenicity
and ecology in the soil environment. Can. J. Microbiol. 42: 881-895.
Edlich, W., Lorenz, G., Lyr, H., Nega, E. and Pommer, E.H. 1989. New aspects on the
infection mechanism of Botrytis cinerea Pers. Neth. J. Plant Pathol. 95: 53-62.
Elad, Y. 1992. The use of antioxidants (free radical scavengers) to control grey mould
(Botrytis cinerea) and white mould (Sclerotinia sclerotiorum) in various crops. Plant
Pathol. 41: 417-426.
Elad, Y. and Volpin, H. 1988. The involvement of ethylene, and calcium in gray mold of
pelargonium, Ruscus, and rose plants. Phytoparasitica 16: 119-132.
Ferrari, S., Vairo, D., Ausubel, F.M., Cervone, F. and De Lorenzo, G. 2003. Tandemly
duplicated Arabidopsis genes that encode polygalacturonase-inhibiting proteins are
regulated coordinately by different signal transduction pathways in response to fungal
infection.
Flaishman, M.A. and Kolattukudy, P.E. (1994) Timing of fungal invasion using host's
ripening hormone as a signal. Proc. Natl. Acad. Sci. USA 91: 6579-6583.
Gentile, A.C. 1954. Carbohydrate metabolism and oxalic acid synthesis by Botrytis
cinerea. Plant Physiol. 29: 257-261.
Germeier, C., Hedke, K. and Von Tiedemann, A. 1994. The use of pH-indicators in
diagnostic media for acid-producing plant pathogens. Z. Pflanzenkrankheiten
Pflanzenschutz 101: 498-507.
Gil-Ad, N.L., Bar-Nun, N. and Mayer, A.M. 2001. The possible function of the glcuan
sheath of Botrytis cinerea: effects on the distribution of enzyme activities. FEMS
Microbiol. Letters 199: 109-113.
Godoy, G., Steadman, J.R., Dickman, M.B. and Dam, R. 1990. Use of mutants to
demonstrate the role of oxalic acid in pathogenicity of Sclerotinia sclerotiorum on
Phaseolus vulgaris. Physiol. Mol. Plant Pathol. 37: 179-191.
Gourgues, M., Brunet-Simon, A., Lebrun, M.H. and Levis, C. 2003. The tetraspanin
BcPls1p is required for appressorium-mediated penetration of Botrytis cinerea into
host plant leaves. Molecular Microbiology, in press.
Govrin, E. and Levine, A. 2000. The hypersensitive response facilitates plant infection by
the necrotrophic pathogen Botrytis cinerea. Current Biol. 10: 751-757.
Hain, R., Reif, H.J., Krause, E., Langebartels, R., Kindl, H., Vornam, B., Wiese, W.,
Schmelzer, E. and Schreier, P.H. 1993. Disease resistance results from foreign
85

phytoalexin expression in a novel plant. Nature 361: 153-156.


Hamada, W., Reignault, P., Bompeix, G. and Boccara, M. 1994. Transformation of
Botrytis cinerea with the hygromycin B resistance gene, hph. Curr. Genet. 26: 251255.
Harper, A.M., Strange, R.N. and Langcake, P. 1981. Characterisation of the nutrients
required by Botrytis cinerea to infect broad bean leaves. Physiol. Plant Pathol. 19:
153-167.
Heale, J.B. and Sharman, S. 1977. Induced resistance to Botrytis cinerea in root slices and
tissue cultures of carrot (Daucus carota L.). Physiol. Plant Pathol. 10: 51-61.
Howard, R.J., Ferrari, M.A., Roach, D.H. and Money, N.P. (1991) Penetration of hard
substrates by a fungus employing enormous turgor pressures. Proc. Natl. Acad. Sci.
USA 88: 11281-11284.
Jarvis, W.R. 1977. Botryotinia and Botrytis species - Taxonomy, physiology and
pathogenicity. A guide to the literature, Monograph no. 14, Ottawa, Research Branch,
Canada Department of Agriculture.
Jeandet, P., Sbaghi, M. and Bessis, R. 1992. The use of phytoalexin induction and of in
vitro methods as a tool for screening grapevines for resistance to Botrytis cinerea. pp.
109-118 in K. Verhoeff, N.E. Malathrakis and B. Williamson (eds.), Recent advances
in Botrytis research, Pudoc Scientific Publishers, Wageningen.
Johnston, D.J. and Williamson, B. 1992. Purification and characterisation of four
polygalacturonases from Botrytis cinerea. Mycol. Res. 96: 343-349.
Kepczynska, E. 1993. Involvement of ethylene in the regulation of growth and
development of the fungus Botrytis cinerea Pers. ex. Fr. Plant Growth Regulation 13:
65-69.
Kepczynski, J. and Kepczynska, E. 1977. Effect of ethylene on germination of fungal
spores causing fruit rot. Fruit Science reports 15: 31-35.
Lamb, C.J. and Dixon, R.A. 1997. The oxidative burst in plant disease resistance. Annual
Rev. Plant Physiol. Plant Mol. Biol. 48: 251-275.
Langcake, P. 1981. Disease resistance of Vitis spp. and the production of the stress
metabolites resveratrol, epsilon -viniferin, alpha - viniferin and pterostilbene. Physiol.
Plant Pathol. 18: 213-226.
Legendre, L., Rueter, S., Heinstein, P.F. and Low, P.S. 1993. Characterisation of the
oligogalacturonide-induced oxidative burst in cultured soybean (Glycine max) cells.
Plant Physiol. 102: 233-240.
Liu, S., Oeljeklaus, S., Gerhardt, B. and Tudzynski, B. 1998. Purification and
characterisation of glucose oxidase of Botrytis cinerea, Physiol. Mol. Plant Pathol. 53:
123-132.
Mansfield, J.W. 1980. Mechanisms of resistance to Botrytis. pp. 181-218 in J.R. ColeySmith, K. Verhoeff and W.R. Jarvis (eds.), The biology of Botrytis, Academic Press,
New York.
Mansfield, J.W. and Richardson, A. 1981. The ultrastructure of interactions between
Botrytis species and broad bean leaves. Physiol. Plant Pathol. 19: 41-48.
Maule, A.J. and Ride, J.P. (1976) Ammonia-lyase and O-methyl transferase activities
related to lignification in wheat leaves infected with Botrytis. Phytochemistry 15:
1661-1664.
McNicol, R.J., Williamson, B. and Young, K. 1989. Ethylene production by black currant
flowers infected by Botrytis cinerea. Acta Hort. 262: 209-215.
Mendgen, K., Hahn, M. and Deising, H. 1996. Morphogenesis and mechanisms of
penetration by plant pathogenic fungi. Annual Rev. Phytopathol. 34: 367-386.
Movahedi, S. and Heale, J.B. 1990. The roles of aspartic proteinase and endo-pectin lyase
enzymes in the primary stages of infection and pathogenesis of various host tissues by
different isolates of Botrytis cinerea. Physiol. Mol. Plant Pathol. 36: 303-324.
Muckenschnabel, I., Goodman, B.A., Deighton, N., Lyon, G.D. and Williamson, B.
2001a. Botrytis cinerea induces the formation of free radicals in fruits of Capsicum
annuum at positions remote from the site of infection. Protoplasma 218: 112-116.
86

Muckenschnabel, I., Williamson, B., Goodman, B.A., Lyon, G.D., Stewart, D. and
Deighton, N. 2001b. Markers for oxidative stress associated with soft rots in French
beans (Phaseolus vulgaris) infected by Botrytis cinerea. Planta 212: 376-381.
Muckenschnabel, I., Goodman, B.A., Williamson, B., Lyon, G.D. and Deighton, N. 2002.
Infection of leaves of Arabidopsis thaliana by Botrytis cinerea: Changes in ascorbic
acid, free radicals and lipid peroxidation products. J. Exp. Botany 53: 207-214.
Muckenschnabel, I., Schulze Gronover, C., Deighton, N., Goodman, B.A., Lyon G.D.,
Stewart D. and Williamson, B. 2003. Oxidative effects in uninfected tissue in leaves
of French bean (Phaseolus vulgaris) containing soft rots caused by Botrytis cinerea. J.
Sci. Food Agric. 83: 507-514.
Osbourn, A.E., Melton, R.E., Wubben, J.P., Flegg, L.M., Oliver, R.P. and Daniels, M.J.
1998. Saponin detoxification and fungal pathogenesis. pp. 309-315 in K. Kohmoto
and O.C. Yoder (eds.), Molecular genetics of host-specific toxins in plant diseases,
Kluwer Academic Publishers, Dordrecht.
Pezet, R. 1998. Purification and characterisation of a 32-kda laccase-like stilbene oxidase
produced by Botrytis cinerea Pers. :Fr. FEMS Microbiology Letters 167: 203-208.
Pezet, R., Pont, V. and Hoang Van, K. 1991. Evidence for oxidative detoxification of
pterostilbene and resveratrol by a laccase-like stilbene oxidase produced by Botrytis
cinerea. Physiol. Mol. Plant Pathol. 39: 441-450.
Powell A.L.T, Van Kan J.A.L., Ten Have A., Visser J., Greve L.C., Bennett A.B. and
Labavitch J.M. 2000. Transgenic expression of pear PGIP in tomato limits fungal
colonization. Mol.Plant-Microbe Interact. 13: 942-950.
Prins, T.W., Wagemakers, C.A.M. and Van Kan, J.A.L. 2000a. Cloning and
characterization of a Glutathione S-Transferase homologue from the plant pathogenic
fungus Botrytis cinerea, Mol Plant Pathol. 1: 169-178
Prins, T.W., Tudzynski, P., Von Tiedemann, A., Tudzynski, B., Ten Have, A., Hansen,
M.E., Tenberge, K. and Van Kan, J.A.L. 2000b. Infection strategies of Botrytis
cinerea and related necrotrophic pathogens. pp.33-64 in J. Kronstad (ed.), Fungal
Pathology, Kluwer Academic Publishers.
Prusky, D. 1996. Pathogen quiescence in post-harvest diseases. Annual Rev. Phytopathol.
34:413-434.
Quidde, T., Osbourn, A.E. and Tudzynski, P. 1998. Detoxification of alpha-tomatine by
Botrytis cinerea. Physiol. Mol. Plant Pathol. 52: 151-165.
Rebordinos, L., Cantoral, J.M., Prieto, M.V., Hanson, J.R. and Collado, I.G. 1996. The
phytotoxic activity of some metabolites of Botrytis cinerea. Phytochemistry 42: 383387.
Reignault, P., Mercier, M., Bompeix, G. and Boccara, M. 1994. Pectin methylesterase
from Botrytis cinerea: Physiological, biochemical and immunochemical studies.
Microbiology 140: 3249-3255.
Salinas, J. and Verhoeff, K. 1995. Microscopical studies of the infection of gerbera
flowers by Botrytis cinerea. Eur. J. Plant Pathol. 101: 377-386.
Salinas, J. C. 1992. Function of cutinolytic enzymes in the infection of gerbera flowers by
Botrytis cinerea, Ph.D.-thesis, University of Utrecht, The Netherlands.
Sbaghi, M., Jeandet, P., Bessis, R. and Leroux, P. 1996. Degradation of stilbene-type
phytoalexins in relation to the pathogenicity of Botrytis cinerea to grapevines. Plant
Pathol. 45: 139-144.
Schoonbeek, H., Del Sorbo, G. and de Waard, M.A. 2001. The ABC transporter BcatrB
affects the sensitivity of Botrytis cinerea to the phytoalexin resveratrol and the
fungicide fenpiclonil. Mol. Plant-Microbe Interact. 14: 562-571.
Schoonbeek, H., Raaijmakers, J. and de Waard, M.A. 2002. Fungal ABC transporters and
microbial interactions in natural environments. Mol. Plant-Microbe Interact. 15: 11651172.
Schouten, A., Tenberge, K.B., Vermeer, J., Stewart, J. Wagemakers, C.A.M., Williamson,
B. and van Kan, J.A.L. 2002a. Functional analysis of an extracellular catalase of
Botrytis cinerea. Mol. Plant Pathol. 3: 227-238.
87

Schouten A., Wagemakers, C.A.M., Stefanato, F., Van der Kaaij R.M. and van Kan,
J.A.L. 2002b. Resveratrol acts as a natural profungicide and induces self-intoxication
by a specific laccase. Mol. Microbiol. 43: 883-894.
Schwacke, R. and Hager, A. (1992) Fungal elicitors induce a transient release of active
oxygen species from cultured spruce cells that is dependent on Ca2+ and proteinkinase activity. Planta 187: 136-141.
Staples, R.C. and Mayer, A.M. 1995. Putative virulence factors of Botrytis cinerea acting
as a wound pathogen. FEMS Microbiol. Letters 134: 1-7.
Tenberge, K.B., Beckedorf, M., Hoppe, B, Schouten, A., Solf, M. and von den Driesch,
M. 2002. In situ localization of AOS in host-pathogen interactions. MIcrosc.
Microanal. 8 (suppl. 2): 250-251.
Ten Have, A., Mulder, W., Visser, J. and van Kan, J.A.L. 1998. The endopolygalacturonase gene Bcpg1 is required for full virulence of Botrytis cinerea. Mol.
Plant-Microbe Interact. 11: 1009-1016.
Ten Have, A., Wubben, J.P., Oude-Breuil, W., Mulder, W., Visser, J. and van Kan, J.A.L.
2001. Botrytis cinerea endopolygalacturonase genes are differentially expressed in
various plant tissues. Fungal Genet. Biol. 33: 97-105.
Ten Have, A., Tenberge, K.B., Benen, J.A.E., Tudzynski, P., Visser, J. and van Kan,
J.A.L. 2002. The contribution of cell wall degrading enzymes to pathogenesis of
fungal plant pathogens. pp. 341-358 in F. Kempken (ed.), The Mycota XI,
Agricultural applications, Springer Verlag, Berlin/Heidelberg/New York.
Thomzik, J.E., Stenzel, K., Stocker, R., Schreier, P.H., Hain, R., and Stahl, D.J. (1997)
Synthesis of a grapevine phytoalexin in transgenic tomatoes (Lycopersicon
esculentum mill.) conditions resistance against Phytophthora infestans. Physiol. Mol.
Plant Pathol.51: 265-278.
Valette-Collet, O., Cimerman, A., Reignault, P., Levis, C. and Boccara, M. 2003.
Disruption of Botrytis cinerea pectin methylesterase gene reduces virulence on several
host plants. Mol. Plant-Microbe Interact. 16: 360-367.
van den Heuvel, J. 1981. Effect of inoculum composition on infection of French bean
leaves by conidia of Botrytis cinerea. Neth. J. Plant Pathol. 87: 55-64.
van den Heuvel, J. and Waterreus, L.P. 1983. Conidial concentration as an important
factor determining the type of infection structures formed by Botrytis cinerea on
leaves of French bean (Phaseolus vulgaris). Plant Pathol. 32: 263-272.
van der Vlugt-Bergmans, C.J.B., Wagemakers, C.A.M. and van Kan, J.A.L. 1997a.
Cloning and expression of the cutinase A gene of Botrytis cinerea. Mol. PlantMicrobe Interact. 10: 21-29.
van der Vlugt-Bergmans, C.J.B., Wagemakers, C.A.M., Dees, D.C.T. and van Kan, J.A.L.
1997b. Catalase A from Botrytis cinerea is not expressed during infection on tomato
leaves. Physiol. Mol. Plant Pathol. 50: 1-15.
van Kan, J.A.L., van 't Klooster, J.W., Wagemakers, C.A.M., Dees, D.C.T. and van der
Vlugt-Bergmans, C.J.B. 1997. Cutinase A of Botrytis cinerea is expressed, but not
essential, during penetration of gerbera and tomato. Mol. Plant-Microbe Interact. 10:
30-38.
Verhoeff, K. and Liem, J.I. 1975. Toxicity of tomatine to Botrytis cinerea, in relation to
latency. Phytopathologische Z. 82: 333-338.
Verhoeff, K., Leeman, M., van Peer, R., Posthuma, L., Schot, N. and van Eijk, G.W.
1988. Changes in pH and the production of organic acids during colonisation of
tomato petioles by Botrytis cinerea. J. Phytopathol. 122: 327-336.
Von Tiedemann, A. 1997. Evidence for a primary role of active oxygen species in
induction of host cell death during infection of leaves with Botrytis cinerea. Physiol.
Mol. Plant Pathol. 50: 151-166.
Weigend, M. and Lyr, H. 1996. The involvement of oxidative stress in the pathogenesis
of Botrytis cinerea on Vicia faba leaves. Z. Pflanzenkrankheiten Pflanzenschutz 103:
310-320.
Williamson, B., Duncan, G.H., Harrison, J.G., Harding, L.A., Elad, Y. and Zimand, G.
88

1995. Effect of humidity on infection of rose petals by dry-inoculated conidia of


Botrytis cinerea. Mycol. Res. 99: 1303-1310.
Wubben, J.P., Mulder, W., Ten Have, A., Van Kan, J.A.L. and Visser, J. 1999. Cloning
and partial characterization of the endopolygalacturonase gene family from Botrytis
cinerea. Appl. Environm. Microbiol. 65: 1596-1602.

Figure

Spore attachment

Germination

Appressorium

Penetration of host surface


Generation of
oxidative stress
Killing of host tissue

Evasion
Primary lesion formation

of host
defence
responses

(Quiescence)

Lesion expansion,
tissue maceration

Sporulation

Fig. 1. Different stages in the infection process of B. cinerea. The shaded box represents
the host tissue.

89

90

Das könnte Ihnen auch gefallen